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Molecular pathology of osteopontin transgenic mice

骨桥蛋白转基因小鼠的分子病理学

基本信息

批准号：
06454194
负责人：
HIGUCHI Yasunori
金额：
$ 4.61万
依托单位：
Oita Medical University
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (B)
财政年份：
1993
资助国家：
日本
起止时间：
1993 至 1995
项目状态：
已结题

项目摘要

1.Establishment of transgenic mice (TM) with metallothionein (MT) promoter-driven mouse osteopontin (OP) gene and their analysis : A MT promotor-OP fusiongene was constructed, and everal founder mice were produced. However, use of the TM was impossible because of few number of litter mates and killing of new born TM by mother. Speciemens and samples from a few founder mice were subjected to biochemical and immunohistochemical analysis. Expression of transgenic RNA was abundant in the liver in TM on the basal diet. After maintenance with water containing ZnSO4 (50mM) for 4 days, expression of transgenic RNA was strong in the small intestine. Immunohistochemical analysis demonstrated OP expression in these organs in TM.Levels of OP in sera from TM after maintenance with water containing ZnSO4 (50mM) for 4 days were significantly higher than these animals maintained with normal water and nontransgenic mice. Founder mice were healthy and one of them is alive 2 years after birth. Autopsy ex … More amination of TM revealed no abnormal organs. 2.Establishment of trnansgenic mice (TM) with cytomegalovirus (CV) or U2alpha globulin (U2alpha) promoter-driven OP gene and their analysis : CV or U2alpha promoter-mouse osteopontin (OP) fusiongene was constructed, and a few founder mice were produced. However, we were anable to expand these TM.Speciemens and samples from a few founder mice were subjected to biochemical and immunohistochemical analysis. Northern blot and PAP analysis demonstrated OP in kidney in CVTM and in the liver in U2alpha TM.Levels of OP in sera from TM were significantly higher than nontransgenic mice. 3.Establishment of TM with alpha1-antitrypsin (alpha1AT) promoter-driven OP gene and their analysis : A alpha1AT-OP fusiongene was constructed, and several founder mice were produced in C57BL/6N/DBA/2F1 mice. Expansion of alpha1AT TM was easy. Backcross of heterozygote mouse to C57BL/6N to get TM of this backgrownd is now underway, reaching 6N.Northern blot and PAP analysis demonstrated OP in the liver in alpha1AT TM.Levels of OP in sera from TM were significantly higher than nontransgenic mice. Less

1.金属硫蛋白（MT）启动子驱动的小鼠骨桥蛋白（OP）基因转基因小鼠（TM）的建立及分析：构建MT启动子-OP融合基因，并建立了多只建立者小鼠。然而，由于同窝仔数量少和母亲杀死新生TM，因此无法使用TM。对来自一些创始小鼠的物种和样品进行生物化学和免疫组织化学分析。在基础日粮中，转基因RNA在TM的肝脏中表达丰富。用含ZnSO_4（50 mM）的水维持4天后，转基因RNA在小肠中的表达强烈。免疫组化分析表明，OP表达在这些器官中TM。OP水平从TM的血清中的水含有硫酸锌（50 mM）的4天后，维护显着高于这些动物保持正常的水和nontransgenic小鼠。创始小鼠是健康的，其中一只在出生后2年还活着。尸检前 ...更多信息 TM胺化未发现异常器官。2.巨细胞病毒（cytomegalovirus，CV）或U2 α球蛋白（U2 alpha globulin，U2 alpha）启动子驱动的OP基因转基因小鼠（transmansgenic mice，TM）的建立及其分析：构建了CV或U2 alpha启动子-小鼠骨桥蛋白（osteopontin，OP）融合基因，并获得了少量的创始小鼠。然而，我们能够扩大这些TM。物种和样品从一些创始人小鼠进行生化和免疫组化分析。北方印迹和PAP分析证实CVTM小鼠肾脏和U2 α TM小鼠肝脏中存在OP。TM小鼠血清中OP水平显著高于非转基因小鼠。3.α 1-抗胰蛋白酶（alpha 1-antitrypsin，alpha 1AT）启动子驱动的OP基因TM的建立及分析：构建了α 1AT-OP融合基因，并在C57 BL/6 N/DBA/2F 1小鼠中产生了数只创始小鼠。alpha 1AT TM的扩展很容易。用C57 BL/6 N小鼠与杂合子小鼠进行回交，获得了转基因小鼠的TM，现已达到6 N。Northern杂交和PAP分析表明，α 1AT TM小鼠肝脏中存在OP，TM小鼠血清中OP水平显著高于非转基因小鼠。少